G cap-stabilized oligonucleotides

ABSTRACT

Oligonucleotides of the formula 
     
         5&#39;-(CAP)-(Oligo)-(CAP)-3&#39; 
    
     are disclosed where (oligo) is a nucleotide sequence of from 10 to 40 nucleotides in length, and CAP is G m , where m is an integer of from zero to ten, the two CAP&#39;s which are present in the molecule can be defined independently of each other and must be different in the case where m is zero at the 5&#39; or 3&#39; end and the end of the Oligo sequence is other than guanine. The oligonucleotides can be synthesized chemically. The oligonucleotides are used to diagnose or treat cancer, restenosis, a disease caused by a virus, a disease affected by integrins or cell-cell adhesion receptor or a disease triggered by diffusible factors.

FIELD

The invention relates to oligonucleotides, particularly G Cap-stabilizedoligonucleotides.

BACKGROUND

Antisense oligonucleotides (AO), triple helix-forming oligonucleotides(TFO) and sense oligonucleotides have been found to be specificinhibitors of gene expression in a large number of systems, both invitro and in vivo (e.g., Uhlmann & Peyman, Chem. Rev., 1990, 90: 543;Milligan et al., J. Med. Chem., 1993, 36: 1923; Stein & Cheng, Science,1993, 261: 1004; Bielinaki et al., Science, 1990, 250: 997).

One of the main problems when using naturally occurring phosphodiester(PO) oligonucleotides is their rapid degradation by various nucleolyticactivities both in cells and in the cell culture medium. A variety ofchemical modifications have been employed in order to stabilizeoligonucleotides. Reviews on the state of the art are provided, forexample, by Uhlmann & Peyman, above, and P. D. Cook (Antisense Researchand Applications, Crooke and Lebleu, Eds., Chapter 9, p. 149ff, CRCPress Boca Raton 1993).

Stabilization against nucleolytic degradation can be effected bymodifying or replacing the phosphate bridge, the sugar structuralcomponent or the nucleotide base, or by replacing the sugar-phosphatebackbone of the oligonucleotides. Numerous modifications of theinternucleoside bridge have, in particular, been described, since thephosphate bridge is the center of the nucleolytic attack. Thenuclease-resistant internucleoside bridges which are most frequentlyused are phosphorothioate (PS), methyl phosphonate (MeP) andphosphoramidate (PA) bridges. Hairpin or self-stabilizedoligonucleotides, as described, for example, in Tang et al., Nucl. AcidsRes., 1993, 21: 2729) represent a further option for stabilization.

An additional problem in antisense technology is that cell penetrationby the oligonucleotides is often inadequate. Numerous chemicalmodifications have been employed to improve this situation as well.Uhlmann & Peymann, above, and P. D. Cook, above, provide reviews of thestate of the art. These modifications involve, inter alia, lipophilicconjugate groups at the 5' or 3' end of the oligonucleotide and neutralor ionic modifications of the backbone, and also 2' modifications on thepentofuranosyl ring. Hughes, Antisense Research and Development, 1994,4: 211, demonstrates that the cell uptake of a 10-mer homo-Gphosphorothioate is higher, by a factor of 2, than the cell uptake ofthe 10-mer homo-oligomeric phosphorothioate of T, A or C.

Blackburne, Nature, 1991, 350: 569, describes structures which can beassumed by G-rich oligonucleotides. In the presence of Na⁺ or K⁺, G-richoligonucleotides which possess at least four short segments containing Gresidues are able to form intramolecular structures which containso-called G quartets as the stabilizing element; these quartets comprisefour guanine residues which are linked in one plane by way of Hoogstenbase pairing. Several such elements are arranged one above the other.Frequently, oligonucleotides are too short to form such intramolecularstructures. In these cases, G quartet structures can be formed by theassociation of two separate oligonucleotides.

SUMMARY

It has now been found that a very simple option exists for significantlyimproving unmodified or modified oligonucleotides with regard to theirnuclease resistance and cell penetration, so that their activity issubstantially improved, by extending the oligonucleotides at the 3'and/or 5' end by from one to 10 guanines.

Surprisingly, the novel oligonucleotides also exhibit a tendency toassociate or aggregate. It is possible that they too form G quartetstructures by the association of two or more oligonucleotides. Suchstructures would protect against exonuclease degradation and lead to anincreased uptake into the cell. Since the associated structures arealways also in equilibrium with the "free" oligonucleotides, sufficient"free" oligonucleotide should also always be available for regulatingtranslation or transcription.

DETAILED DESCRIPTION

The invention relates to oligonucleotides of the formula:

    5'-(CAP)-(Oligo)-(CAP)-3'                                                     where Oligo is, for                                                           example (SEQ ID NO:1-34, respectively),                                       ACACCCAATTCTGAAAATGG,     (I)                                                 AGGTCCCTGTTCGGGCGCCA,     (II)                                                GTCGACACCCAATTCTGAAAATGGATAA,                                                                           (III)                                               GCTATGTCGACACCCAATTCTGAAA,                                                                              (IV)                                                GTCGCTGTCTCCGCTTCTTCTTCCTG,                                                                             (V)                                                 GTCTCCGCTTCTTCTTCCTGCCATAGG,                                                                            (VI)                                                GCGGGGCTCCATGGGGGTCG,     (VII)                                               CAGCTGCAACCCAGC,          (VIII)                                              GGCTGCTGGAGCGGGGCACAC,    (IX)                                                AACGTTGAGGGGCAT,          (X)                                                 GTGCCGGGGTCTTCGGGC,       (XI)                                                GGAGAACATCATGGTCGAAAG,    (XII)                                               CCCGAGAACATCATGGTCGAAG,   (XIII)                                              GGGGAAAGCCCGGCAAGGGG,     (XIV)                                               CACCCGCCTTGGCCTCCCAC,     (XV)                                                GGGACTCCGGCGCAGCGC,       (XVI)                                               GGCAAACTTTCTTTTCCTCC,     (XVII)                                              GGGAAGGAGGAGGATGAGG,      (XVIII)                                             GGCAGTCATCCAGCTTCGGAG,    (XIX)                                               GCAGTAAGCATCCATATC,       (XX)                                                CCCCCACCACTTCCCCTCTC,     (XXI)                                               CTCCCCCACCACTTCCCCTC,     (XXII)                                              GCTGGGAGCCATAGCGAGG,      (XXIII)                                             ACTGCTGCCTCTTGTCTCAGG,    (XXIV)                                              CAATCAATGACTTCAAGAGTTC,   (XXV)                                               GGTCCCTGTTCGGGCGCCA,      (XXVI)                                              GTGCCGGGGTCTTCGGG,        (XXVII)                                             GGAGGATGCTGAGGAGG,        (XXVIII)                                            GGAGGATGCTGAGG,           (XXIX)                                              CAGGAGGATGCTGAGGAGG,      (XXX)                                               GGCTGCCATGGTCCC,          (XXXI)                                              TCATGGTGTCCTTTGCAGCC,     (XXXII)                                             TCATGGTGTCCTTTGCAG, or    (XXXIII)                                            AAGTTCATGGTTTCGG,         (XXXIV)                                         

and CAP is G_(m), where m is an integer of from zero to ten, preferablyof from two to six, more preferably of from three to five and still morepreferably four, and where the two CAP's which are present in themolecule can be defined independently of each other and must bedifferent in the case where m is zero at the 5' or 3' end, wherein theend of the "Oligo" sequence is other than guanine.

In those cases in which the oligonucleotide (Oligo) ends at the 5' or 3'end with one or more guanines, it can be advantageous if CAP isG(_(m-n)), where m in defined as above and n is the number of theguanines which naturally occur at the 5' or 3' end of theoligonucleotide (Oligo), and where (m-n) is preferably from two to six,more preferably from three to five, and still more preferably four.

The oligonucleotides which have been adapted in this way can beunmodified or modified, with the following variants being permitted:

a) Complete or partial replacement of the 3' and/or 5' phosphoricdiester bridges, for example by a phosphorothioate, phosphorodithioate,(NR¹ R²)-phosphoramidate, boranophosphate, phosphate (C₁ -C₂₁)-O-alkylester, phosphate [(C₆ -C₁₂) aryl-(C₁ -C₂₁)-O-alkyl] ester,2,2,2-trichlorodimethylethylphosphonate, (C₁ -C₈) alkylphosphonate or(C₆ -C₁₂)arylphosphonate bridge.

Replacement by a phosphorothioate, phosphorodithioate, NR¹ R²-phosphoramidate, phosphate O-methyl ester, phosphate O-ethyl ester,phosphate O-isopropyl ester, methylphosphonate or phenylphosphonatebridge in preferred. Replacement by a phosphorothioate,phosphorodithioate or methylphosphonate bridge is more preferred.Replacement by a phosphorothioate bridge is still more preferred.

R¹ and R² are, independently of each other, hydrogen or (C₁ -C₁₈)-alkyl,(C₆ -C₂₀)-aryl, (C₆ -C₁₄)-aryl-(C₁ -C₈)-alkyl or --(CH₂)_(c)--(NH(CH₂)_(c))_(d) -NR³ R³, in which c is an integer of from 2 to 6,and d is an integer of from 0 to 6, and the R³ groups are, independentlyof each other, hydrogen, (C₁ -C₆)-alkyl or (C₁ -C₄)-alkoxy-(C₁-C₆)-alkyl; R¹ and R² are preferably hydrogen, (C₁ -C₈)-alkyl ormethoxyethyl, more preferably hydrogen, (C₁ -C₄)-alkyl or methoxyethyl.R¹ and R² can also, together with the nitrogen atom carrying them, forma 5-6-membered heterocyclic ring which can additionally contain afurther heteroatom from the series O, S and N.

Preferably, one, two or three phosphoric diester bridges should bereplaced at the 5' end and/or at the 3' end, preferably at the 5' endand at the 3' end. Preferably, the replacement of the phosphoric diesterbridges should also be effected at the pyrimidine positions.

b) Complete or partial replacement of the 3' or 5' phosphoric diesterbridges by "dephospho" bridges (see, for example, Uhlmann and Peyman inMethods in Molecular Biology, vol. 20: "Protocols for Oligonucleotidesand Analogs", S. Agrawal, Ed., Humana Press, Totowa 1993, Chapter 16,355ff), for example by formacetal, 3'-thio-formacetal,methylhydroxylamine, oxime, methylenedimethylhydrazo, dimethylenesulfoneor silyl groups. Replacement by formacetals and 3'-thioformacetals ispreferred.

Preferably, one, two or three phosphoric diester bridges should bereplaced at the 5' end and/or at the 3' end, preferably at the 5' endand at the 3' end. Preferably, the replacement of the phosphoric diesterbridges should also be effected at the pyrimidine positions.

c) Complete or partial replacement of the sugar phosphate backbone, forexample by "morpholinonucleoside" oligomers (see E. P. Stirchak et al.,Nucleic Acids Res., 1989, 17: 6129) or peptide nucleic acids (PNA's)(see P. E. Nielsen et al., Bioconj. Chem., 1994, 5: 3) or else PNA/DNAhybrids as described in German Patent Application P 44 08 528.1.

d) Complete or partial replacement of the β-D-2'-deoxyribose units, forexample by α-D-2'-deoxyribose, L-2'-deoxyribose, 2'-F-2'-deoxyribose,2'-O-(C₁ -C₆)alkyl ribose, 2'-O-(C₂ -C₆)alkenyl ribose, 2'-NH₂-2'-deoxyribose, β-D-xylofuranose, α-arabinofuranose,2,4-dideoxy-β-D-erythrohexopyranose, and carbocyclic (see Froehler, J.Am. Chem. Soc., 1992, 114: 8320) and open-chain sugar analogs (seeVandendriesoche et al., Tetrahedron, 1993, 49: 7223) and bicyclo sugaranalogs (see M. Tarkov et al., Helv. Chim. Acta., 1993, 76: 481).

Replacement by 2'-F-2'-deoxyribose, 2'-O-(C₁ -C₆) alkyl ribose, 2'-O-(C₂-C₆) alkenyl ribose or 2'-NH₂ -2'-deoxyribose is preferred. Replacementby 2'-F-2'-deoxyribose, 2'-O-(C₁ -C₄) alkyl ribose, 2'-O-(C₂ -C₄)alkenyl ribose or 2'-NH₂ -2'-deoxyribose is more preferred. Replacementby 2'-O-methylribose, 2'-O-allylribose or 2'-O-butylribose is still morepreferred.

Preferably, one, two or three ribose units should be replaced at the 5'end and/or at the 3' end, preferably at the 5' end and at the 3' end.Preferably, the replacement of the ribose units should also be effectedat the pyrimidine positions.

e) Complete or partial replacement of the natural nucleoside bases, forexample by 5-(hydroxymethyl)uracil, 5-aminouracil, pseudouracil,dihydrouracil, 5-(C₁ -C₆)-alkyl-uracil, 5-(C₂ -C₆)-alkenyl-uracil, 5-(C₂-C₆)-alkynyl-uracil, 5-(C₁ -C₆)-alkyl-cytosine, 5-(C₂-C₆)-alkenyl-cytosine, 5-(C₂ -C₆)-alkynyl-cytosine, 5-fluorouracil,5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil,5-bromocytosine or 7-deaza-7-substituted purines.

Replacement by 5-(C₁ -C₆)-alkyl-uracil, 5-(C₂ -C₆)-alkenyluracil, 5-(C₂-C₆)-alkynyl-uracil, 5-(C₁ -C₆)-alkylcytosine, 5-(C₂-C₆)-alkenyl-cytosine, 5-(C₂ -C₆)-alkynylcytosine, 5-fluorouracil,5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil,5-bromocytosine, 7-deaza-7-alkynyl, preferably hexynyl-substitutedpurines, 7-deaza-7-methyl-substituted purines or7-deaza-7-bromine-substituted purines is preferred. Replacement by 5-(C₃-C₆)-alkyl-uracil, 5-(C₂ -C₆)-alkenyluracil, 5-(C₂ -C₆)-alkynyl-uracil,5-(C₁ -C₆)-alkylcytosine, 5-(C₂ -C₆)-alkenyl-cytosine or 5-(C₂-C₆)-alkynylcytosine is more preferred. Replacement by5-hexynylcytosine, 5-hexynyluracil, 5-hexynylcytosine, 5-propynyluracilor 5-propynylcytosine is still more preferred.

The nucleoside bases should not be replaced in the CAP regions.

Of the above-mentioned modifications, those given in groups a), b), c)and d) are especially preferred. More preferred are those modificationsgiven in groups a) and d), with those given in group a) being mostpreferred.

In addition, the novel oligonucleotides can be linked to (or conjugatedwith), for example at the 3' or 5' end, molecules which are known tohave a favorable influence on the properties of antisenseoligonucleotides or triple helix-forming oligonucleotides (such as, forexample, cell penetration, nuclease degradation, affinity for the targetRNA/DNA, and pharmacokinetics). Examples are conjugates with polylysine,with intercalators such as pyrene, acridine, phenazine andphenanthridine, with fluorescent compounds such as fluorescein, withcross-linkers such as psoralene and azidoproflavin, with lipophilicmolecules such as (C₁₂ -C₂₀)-alkyl, with lipids such as1,2-dihexadecyl-rac-glycerol, with steroids such as cholesterol ortestosterone, with vitamins such an vitamin E, with polyethylene glycolor oligoethylene glycol, with (C₁₂ -C₁₈)-alkyl-phosphate diesters, andwith --O--CH₂ --CH(OH)-O-(C₁₂ -C₁₈)-alkyl. Conjugates with lipophilicmolecules, such as (Cl₂ -C₂₀)-alkyl, with steroids, such as cholesterolor testosterone, with polyethylene glycol or oligoethylene glycol, withvitamin E, with intercalators, such as pyrene, with (C₁₄-C₁₈)-alkyl-phosphate diesters and with --O--CH₂ --CH(OH)--O--(C₁₂-C₆)-alkyl are preferred. Those skilled in the art are familiar with thepreparation of such oligonucleotide conjugates (see, for example,Uhlmann & Peyman, Chem. Rev., 1990, 90: 543; M. Manoharan in AntisenseResearch and Applications, Crooke and Lableu, Eds., CRC Press, BocaRaton, 1993, Chapter 17, p. 303ff; and EP 0552766A2).

Furthermore, the novel oligonucleotides can carry 3'--3'and 5'--5'inversions at the 3' and/or the 5' end (described, for example, in M.Koga et al., J. Org. Chem., 1991 56: 3757).

The invention also relates to a process for preparing the novelcompounds using methods, in particular chemical synthesis, theindividual and separate steps of which are known to a person skilled inthe art, as well as to the use of the novel compounds for preparing apharmaceutical and also to a process for preparing a pharmaceuticalwhich comprises mixing the novel oligonucleotides with a physiologicallyacceptable excipient and, where appropriate, suitable additives and/orauxiliary substances.

In a quite general manner, the present invention also extends to the useof therapeutically effective oligonucleotides, in which at least onenon-terminal pyrimidine nucleoside is modified, for preparing thepharmaceutical. In general, therapeutically effective oligonucleotidesare understood to mean antisense oligonucleotides, triple helix-formingoligonucleotides, aptamers (RNA or DNA molecules which are able to bindto specific target molecules, for example proteins or receptors, e.g.,L.C. Bock et al., Nature, 1992, 355: 564, or ribozymes (catalytic RNA),e.g., Castanetto et al., Critical Rev. Eukar. Gene Expr., 1992, 2: 331),in particular antisense oligonucleotides.

In addition to this, the present invention also relates to the use ofoligonucleotides possessing at least one terminal and modifiedpyrimidine nucleoside as diagnostic agents, for example for detectingthe presence or absence, or the quantity, of a specific double-strandedor single-stranded nucleic acid molecule in a biological sample.

For use according to the invention, the oligonucleotides have a lengthof from about 6 to 60, preferably of from about 10 to 40, in particularof from about 12 to 31, nucleotides. Otherwise, the above-describedpreference ranges, modifications and/or conjugations also apply here.

The pharmaceuticals of the present invention may be used, for example,for treating diseases which are caused by viruses, for example by HIV,HSV-1, HSV-2, influenza, VSV, hepatitis B or papilloma viruses.

Examples of novel antisense oligonucleotides which are effective againstthese targets are:

    a) against HIV, e.g.                                                          5'-G*G*G*G A C A C C C A A T T C T G A A A A T G*G*G*G-3' or                                                             (SEQ ID NO:35)                     5'-G*G*G*G A C A C C*C A A T*T C*T G A A A A T G*G*G*G-3' or                  5'-G*G*G G A C A C*C*C A A T T C T G A A A A T G*G*G*G-3'                     5'-G*G*G*A G G T*C C*C*T G T*T*C G G G C G C*C A G*G*G*G-3' or                                                           (SEQ ID NO:36)                     5'-G*G*G*A G G T*C C*C*T G*T T*C G G G C G C*C*A*G*G*G*G-3'                   5'-G*G*G*G T*C C*C*T G T*T*C G G G C G C*C*A*G*G*G*G-3'                                                                  (SEQ ID NO:37)                     5'-G*G*G G T*C G A*C A C*C C A A T*T C*T G A A A A T*GGA T*A*A-3'                                                        (SEQ ID NOS:38-39)                 5'-G*G*G G T*C G A*C A C*C*C A A T*T C*T G A A A A T*G G A T*A*A-3' or        5'-G*G*G G T*C G A*C A C*C*C A A T*T C*T G A A A A T*G G A*T*A*A-3' or        5'-G*G*G T*C*G A*C A C*C*C A A T*T*C*T G A A A A T*G G A*T*A*A-3'             5'-G*C*T A T G T*C G A*C A C C*C A A T*T*C*T*G A A A G*G*G*G-3' or                                                       (SEQ ID NOS:40-41)                 5'-G*C*T A T*G T*C G A*C A C C*C A A T*T*C*T*G A A A G*G*G*G-3' or            5'-G*C*T A T*G T*C G A C A C*C C*A A T*T C*T G A A A G*G*G*G-3' or            5'-G*C*T A T*G T*C G A C*A C*C C*A A T*T C*T G A A A G*G*G*G-3' or            5'-G*C*T A T G T*C G A C A C*C C*A A T*T C*T G A A A G*G*G-3'                 5'-G*T*C G C*T G T C*T*C*C G C T*T C T T C T T C C*T G*G*G*G-3' or                                                       (SEQ ID NOS:42-44)                 5'-G*T*C G C*T G T C*T*C*C G C T*T C T T C T T C C*T G G*G*G*G-3' or          5'-G*T*C G C*T G T C*T*C*C G C T*T C T T C T T C C*T G G*G*G*G*G-3' or        5'-G*T*C*T C*C G C T*T C*T T*C T*T C*C T G C*C A T A G G*G*G*G-3'                                                        (SEQ ID NOS:45-46)                 5'-G*T*C*T C*C G C T*T C*T T*C T*T C*C T G C*C A T A G*G*G*G-3'or             b) against HSV-1, e.g.                                                        5'-G*C*G G G G C T C C*A T G G G G G T*C*G*G*G-3' or                                                                     (SEQ ID NOS:47-48)                 5'-G*G*C*G G G G C*T C C A*T G G G G G T*C*G-3' or                            5'-G*G*G*G A G G A T*G C*T*G A G G A G G*G*G*G-3' or                                                                     (SEQ ID NOS:49-50)                 5'-G*G*G*G A G G A T*G C*T*G A G G A G G*G*G-3' or                            5'-G*G*G*G G A G G A T*G C*T G A G G*G*G*G-3' or                                                                         (SEQ ID NOS:51-52)                 5'-G*G*G G A G G A T*G C*T G A G G*G*G*G-3' or                                5'-G*G*G*C A G G A G G A T*G C*T*G A G G A G G*G*G*G-3' or                                                               (SEQ ID NOS:53-54)                 5'-G*G*G*G*C A G G A G G A T*G C*T*G A G G A G G*G*G*G-3'.                

The arabic numbers which are given here refer to the Roman numbers givenearlier; the oligonucleotides which are listed above are additionallyprovided with the novel CAP'S.

In the above sequences, the phosphoric diester bonds which were replacedby a phosphorothioate bridge (P=S) were labeled with an asterisk (*).

The pharmaceuticals of the present invention are also suitable, forexample, for treating cancer or restenosis. For example, oligonucleotidesequences may be used in this context which are directed against targetswhich are responsible for the genesis or growth of cancer.

Examples of these targets are:

1) Nuclear oncoproteins such as, for example, c-myc, N-myc, c-myb,c-fos, c-fos/jun, PCNA and p120;

2) Cytoplasmic/membrane-associated oncoproteins such as, for example,EJ-ras, c-Ha-ras, N-ras, rrg, bcl-2, cdc-2, c-raf-1, c-mos, c-src andc-abl;

3) Cellular receptors such as, for example, EGF receptor, c-erbA,retinoid receptors, protein kinase regulatory subunit and c-fms; and

4) Cytokines, growth factors and extracellular matrix such as, forexample, CSF-1, IL-6, IL-1a, IL-1b, IL-2, IL-4, bFGF, myeloblastin,fibronectin and VEGF (vascular endothelial growth factor).

Examples of novel antisense oligonucleotides which are effective againstthese targets are:

    a) against c-Ha-ras, e.g.                                                     5'-G*G*G*G C A C*T G*C A A C*C*C A G*C G*G*G*G-3' or                                                                (SEQ ID NOS:55-56)                      5'-G*G*G*C A G C*T G*C A A C*C*C A G*C G*G*G*G-3' or                          5'-G*G*G*G*C*A*G*C*T*G*C*A*A*C*C*C*A*G*C*G*G*G*G-3' or                        b) c-myc, e.g.                                                                5'-G*G*G*G C*T G C*T G G A G*C G G G G*C A C*A*C-3' or                                                              (SEQ ID NOS:57-58)                      5'-G*G*G*G C*T G C*T G G A G*C G G G G*C A C*A*C*G*G*G*G-3' or                5'-G*G*G*G*C*T*G*C*T*G*G*A*G*C*G*G*G*G*C*A*C*A*C-3' or                        5'-G*G*G*G A A*C G T*T G A G G G G C*A*T-3' or                                                                      (SEQ ID NOS:59-60)                      5'-G*G*G*G A A*C G T*T G A G G G G C*A*T G*G*G*G-3' or                        c) c-myb, e.g.                                                                5'-G*G*G*G T*G C*C G G G G T*C*T*T C G G*G*C-3' or                                                                  (SEQ ID NOS:61-62)                      5'-G*G*G*G T*G C*C G G G G T*C*T*T C G G*G*C G*G*G*G-3' or                    5'-G*G*G*G T*G C*C*G G G G T*C T*T*C G G*G*G*G-3' or                                                                (SEQ ID NO:63)                          d) c-fos, e.g.                                                                5'-G*G*G*G G A G A A C*A T*C A T*G G T*C G A A*A*G-3' or                                                            (SEQ ID NOS:64-67)                      5'-G*G*G*G G A G A A C*A T*C A T*G G T*C G A A A G*G*G*G-3' or                5'-G*G*G*G A G A A C*A T*C A T*G G T*C G A A A G*G*G*G-3' or                  5'-G*G*A G A A C*A*T*C A T*G G T*C G A A*A*G*G*G*G*G-3' or                    5'-C*C*C*G A G A A*C A T*C A T*G G T*C G A*A*G*G*G*G*G-3' or                                                        (SEQ ID NO:68)                          5'-G*G*G G A A A G C*C*C G G*C A A G G*G*G*G-3' or                                                                  (SEQ ID NOS:69-70)                      5'-G*G*G*G G A A A G C*C C*G G C*A A G G*G*G*G-3'                             e) p120, e.g.                                                                 5'-C*A*C*C C*G C*C T*T G G C C T*C C*C A*C G G*G*G*G-3' or                                                          (SEQ ID NOS:71-72)                      5'-C*A*C*C C*G C*C T*T G G C*C T*C C*C A*C G G*G*G-3' or                      f) EGF receptor, e.g.                                                         5'-G*G*G*G A C*T*C*C G G*C G*C A G C*G*C-3' or                                                                      (SEQ ID NOS:73-75)                      5'-G*G*G*G A C*T*C*C G G*C G*C A G C*G*C G*G*G*G-3' or                        5'-G*G*G G G A C*T*C*C G G*C G*C A G C*G*C G G*G*G-3' or                      5'-G*G*G*G C A A A C T*T*T C T T*T*T C C T*C*C-3' or                                                                (SEQ ID NOS:76-77)                      5'-G*G*G*G C A A A C T*T*T C T T*T*T C C T*C*C G G*G*G-3' or                  g) p53 tumor suppressor, e.g.                                                 5'-G*G*G G G A A G G A G G A G G A T*G A*G*G-3' or                                                                  (SEQ ID NOS:78-79)                      5'-G*G*G G G A A G G A G G A G G A T*G A G G G*G*G-3' or                      5'-G*G*G*G*C A G T*C A T*C*C A G C*T T*C G G*A*G-3' or                                                              (SEQ ID NOS:80-81)                      5'-G*G*G G*C A G T*C A T*C*C A G C*T T*C G G A G*G*G*G-3' or                  h) bFGF, e.g.                                                                 5'-G*G*G G C*T G C C A*T G G T*C*C*C-3' or                                                                          (SEQ ID NOS:82-83)                      5-G*G*G*G C*T G C C A*T G G T*C C*C G*G*G*G-3' or                             i) VEGF, e.g.                                                                 5'-G*G*G*G A A G T*T*C A*T G G T*T*T C G*G*G*G-3'.                                                                  (SEQ ID NO:84)                      

The pharmaceuticals of the present invention are furthermore suitable,for example, for treating diseases which are influenced by integrins orcell-cell adhesion receptors, for example by VLA-4, VLA-2, ICAM or ELAM.

Examples of novel antisense oligonucleotides which are effective againstthese targets are:

    a) VLA-4, e.g.                                                                5'-G*G*G*G C*A G*T A A G C*A T*C*C A T*A*T*C-3' or                                                                   (SEQ ID NOS:85-86)                     5'-G*G*G*G C*A G*T A A G C*A T*C*C A T*A T*C G*G*G*G-3' or                    b) ICAM, e.g.                                                                 5'-G*G*G*G*C*C*C C C A C*C A C T*T*C*C C C T C*T*C-3' or                                                             (SEQ ID NOS:87-89)                     5'-C*C*C*C C A C*C A C T*T*C*C C C T*C*T*C*G*G*G*G-3' or                      5'-G*G*G*G*C*C*C*C C A C*C A C T*T*C*C C C T*C*T*C*G*G*G*G-3' or              5'-G*G*G*C*T*C*C C C C A C*C A C T*T C C C*C*T*C*G*G*G*G-3' or                                                       (SEQ ID NOS:90-91)                     5'-G*G*C*T*C*C C C C A C*C A C T*T C C C*C*T*C*G*G*G*G-3' or                  5'-G*G*G*G*C*T G G G A G C*C A*T A G*C G A*G*G-3' or                                                                 (SEQ ID NOS:92-94)                     5'-G*G*G*G*C*T G G G A G C*C A*T A G*C G A*G*G*G*G-3' or                      5'-G*G*G G*C*T G G G A G*C*C A*T A G*C G A G G*G*G*G*3' or                    c) ELAM-1, e.g.                                                               5'-A*C*T G C*T G C*C T*C T*T G T*C T*C A*G*G*G*G-3' or                                                               (SEQ ID NOS:95-96)                     5'-G*G*G*G A C*T G C*T G C*C T*C T*T G T*C T*C A G G*G*G*3' or                5'-G*G*G*G C*A A T*C A A T*G A C*T T*C A A G A G T*T*C-3' or                                                         (SEQ ID NOS:97-98)                     5'-C*A*A T C A A T*G A C*T T*C A A G A G T*T*C G G*G*G-3'                 

The pharmaceuticals of the present invention are also suitable, forexample, for treating diseases which are triggered by factors such asTNF-alpha.

Examples of novel antisense oligonucleotides which are effective againstthese targets are:

    a) TNF-alpha, e.g.                                                            5'-G*G*G*G T C A T G G*T G T C*C T*T T G C A*G*C*C-3' or                                                               (SEQ ID NOS:99-101)                  5'-G*G*G*G T*C A*T G G*T G T C*C T*T*T G*C A G C*C G*G*G*G-3' or              5'-G*G*G*G T*C A*T G G*T G T C*C T*T*T G*C A G C*C G G*G*G*G-3' or            5'-G*G*G*G T*C A*T G G*T G T C*C T*T*T G*C A G G*G*G*G-3' or                                                           (SEQ ID NOS:102-103)                 5'-T*C*A*T G G*T G*T C*C T*T*T G*C A G G*G*G*G-3'                         

That which has been stated above applies with regard to the numbering ofthe exemplary oligonucleotides and the asterisk symbol. The noveloligonucleotides can also be used to prepare one diagnostic agent atleast for all the diseases mentioned.

The pharmaceuticals may be used, for example, in the form ofpharmaceutical preparations which can be administered orally, forexample in the form of tablets, coated tablets, hard or soft gelatincapsules, solutions, emulsions or suspensions. They may also beadministered rectally, for example in the form of suppositories, orparenterally, for example in the form of injection solutions. Forproducing pharmaceutical preparations, these compounds can be workedinto therapeutically inert, organic and inorganic excipients. Examplesof such excipients for tablets, coated tablets and hard gelatin capsulesare lactose, corn starch or derivatives thereof, tallow and stearic acidor salts thereof. Suitable excipients for preparing solutions are water,polyols, sucrose, invert sugar and glucose. Suitable excipients forinjection solutions are water, alcohols, polyols, glycerol and vegetableoils. Suitable excipients for suppositories are vegetable and hardenedoils, waxes, fats and semiliquid polyols.

The pharmaceutical preparations can also contain preservatives,solvents, stabilizers, wetting agents, emulsifiers, sweeteners, dyes,flavorants, salts for altering the osmotic pressure, buffers, coatingagents, antioxidants and, where appropriate, other therapeuticallyactive compounds.

Oral administration and injections are preferred. For injection, theantisense oligonucleotides are formulated in a liquid solution,preferably in a physiologically acceptable buffer such as, for example,Hank's solution or Ringer's solution. However, the antisenseoligonucleotides can also be formulated in solid form and dissolved orsuspended before use. The doses which are preferred for the systemicadministration are from about 0.01 mg/kg to about 50 mg/kg of bodyweight and per day.

The invention is further illustrated by, though in no way limited to,the following examples. Table 1 shows oligonucleotides which have beentested for their in vitro activity against HSV-1. Oligonucleotide no. 4is modified as described in Mann et al. (Bioconj. Chem., 1992, 3: 554)by the introduction of a (4-(1-pyrenyl)butanyl)phosphodiester at the 5'end. The novel oligonucleotides exert an effect at a minimum inhibitoryconcentration which is as low as 9 μM (Examples 1 and 4) or even 3 μM(Examples 2 and 3).

EXAMPLE 1

Oligonucleotide Synthesis

Unmodified oligonucleotides were synthesized on an automatic DNAsynthesizer (Applied Biosystems, model 380B or 394) using standardphosphoramidite chemistry and oxidizing with iodine. For introducingphosphorothioate bridges in mixed phosphorothioates and phosphodiesteroligonucleotides, oxidation was carried out with TETD (tetraethylthiuramdisulfide) instead of with iodine (Applied Biosystems User Bulletin 65).Following cleavage from the solid support (CPG or Tentagel) and removalof the protective groups with conc. NH₃ at 55° C. for 18 h, theoligonucleotides were initially purified by butanol precipitation(Sawadogo, Van Dyke, Nucl. Acids Res., 1991, 19: 674). The sodium saltwas then obtained by precipitating from a 0.5 M NaCl solution containing2.5 parts by volume of ethanol.

The oligonucleotides were analyzed by

a) analytical gel electrophoresis in 20% acrylamide, 8 M urea, 45 mMTris-borate buffer, pH 7.0, and/or

b) HPLC analysis: Waters GenPak FAX, gradient CH₃ CN (400 ml), H₂ O (1.6l), NaH₂ PO₄ (3.1 g), NaCl (11.7 g), pH 6.8 (0.1 M with respect to NaCl)after CH₃ CN (400 ml), H₂ O (1.6 l), NaH₂ PO₄ (3.1 g), NaCl (175.3 g),pH 6.8 (1.5 M with respect to NaCl), and/or

c) capillary gel electrophoresis, Beckman capillary tube eCAP™, U100Pgel column, 65 cm length, 100 mm I.D., window 15 cm from one end, buffer140 μM Tris, 360 mM boric acid, 7M urea, and/or

d) electrospray mass spectroscopy.

The analysis of the oligonucleotides indicated that each of them wasmore than 90% pure.

EXAMPLE 2

Investigation of the in-vitro antiviral activity of test substancesagainst herpes viruses

The antiviral activity of the test substances against various herpesviruses which are pathogenic to humans is investigated in a cell culturetest system. For the experiment, monkey kidney cells (Vero, 2×10⁵ /ml)in serum-containing Dulbecco's MEM (5% fetal calf serum (FCS)) are sownin 96-well microliter plates, which are incubated at 37° C. for 24 h in5% CO₂. The serum-containing medium is then sucked off and the cells arerinsed twice with serum-free Dulbecco's MEM.

The test substances are prediluted in H₂ O to a concentration of 600 μM,and the solutions are stored at -18° C. For the test, further dilutionsteps are carried out in Dulbecco's minimal essential medium (MEM). 100μl of each of the individual test substance dilutions are added,together with 100 μl of serum-free Dulbecco's MEM (-FCS), to the rinsedcells. After 3 h of incubation at 37° C. and in 5% CO₂, the cells areinfected with herpes simplex virus type 1 (ATCC VR733, HSV-1 F strain)or with herpes simplex virus type 2 (ATCC VR734, HSV-2 G strain) inconcentrations at which the cell lawn is completely destroyed within 3days. In the case of HSV-1, the infection intensity is 500plaque-forming units (PFU) per well, while in the case of HSV-2 it is350 PFU/well. The experimental mixtures then contain test substance atconcentrations of from 80 μM to 0.04 μM in MEM, supplemented with 100U/ml penicillin G and 100 mg/l streptomycin. All the experiments arecarried out as duplicate determinations apart from the controls, whichare carried out eight times per plate.

The experimental mixtures are incubated at 37° C. for 17 h in 5% CO₂.The cytotoxicity of the test substances is determined after a totalincubation time of 20 h by microscopic assessment of the cell cultures.The highest preparation concentration which still does not elicit anymicroscopically recognizable cell damage under the stated experimentalconditions is designated the maximum tolerated dose (MTD).

After this, FCS is added to a final concentration of 4% and the platesare incubated for a further 55 h at 37° C. in 5% CO₂. The untreatedinfection controls then exhibit a fully developed cytopathic effect(CPE). After the cell cultures have been assessed microscopically, theyare then stained with neutral red in accordance with the vital stainingmethod of Pinter (1966). The antiviral activity of a test substance isdefined as the minimum inhibitory concentration (MIC) which is requiredin order to protect 30-60% of the cells from the cytopathogenic effectdue to virus.

Table 1 shows the activity of variously modified antisenseoligonucleotides against HSV-1 in cell culture. The phosphodiester bondswhich were replaced by a phosphorothioate bridge (P=S) were labeled withan * in the sequences; MIC=minimum inhibitory concentration; MTD=maximumtolerated dose; Py=pyrene.

                                      TABLE 1                                     __________________________________________________________________________    Sequence                    MIC                                                                              MTD                                            __________________________________________________________________________    5'-G*G*G*C A G G A G G A T*G C*T*G A G G A G G*G*G*G                                                      9  >80 (SEQ ID NO:53)                             5'-G*G*G*G G A G G A T*G C*T*G A G G A G G*G*G*G                                                          3  >80 (SEQ ID NO:104)                            5'-G*G*G*G G A G G A T*G C*T G A G G*G*G*G                                                                3  >80 (SEQ ID NO:51)                             5'-Py-G*G*G*G G A G G A T*G C*T G A G G*G*G*G                                                             9  >80 (SEQ ID NO:105)                            __________________________________________________________________________

It will be apparent to those skilled in the art that variousmodifications and variations can be made to the compositions andprocesses of this invention. Thus, it is intended that the presentinvention cover such modifications and variations, provided they comewithin the scope of the appended claims and their equivalents.

The disclosure of all publications cited above are expresslyincorporated herein by reference in their entireties to the same extentas if each were incorporated by reference individually. The disclosureof German Patent Application No. 195 02 912.7, for which benefit under35 USC § 119 is claimed, is expressly incorporated herein in itsentirety.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 105                                           - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 # 20               ATGG                                                       - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 # 20               GCCA                                                       - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 28 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 #             28   GAAA ATGGATAA                                              - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 #               25 ATTC TGAAA                                                 - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 #              26  CTTC TTCCTG                                                - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 #             27   CCTG CCATAGG                                               - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 # 20               GTCG                                                       - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 15 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 #    15                                                                       - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 #21                CACA C                                                     - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 15 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                #    15                                                                       - (2) INFORMATION FOR SEQ ID NO:11:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 18 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                #  18              GC                                                         - (2) INFORMATION FOR SEQ ID NO:12:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                #21                GAAA G                                                     - (2) INFORMATION FOR SEQ ID NO:13:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                #                 22CGA AG                                                    - (2) INFORMATION FOR SEQ ID NO:14:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                # 20               GGGG                                                       - (2) INFORMATION FOR SEQ ID NO:15:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                # 20               CCAC                                                       - (2) INFORMATION FOR SEQ ID NO:16:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 18 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                #  18              GC                                                         - (2) INFORMATION FOR SEQ ID NO:17:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                # 20               CTCC                                                       - (2) INFORMATION FOR SEQ ID NO:18:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                # 19               AGG                                                        - (2) INFORMATION FOR SEQ ID NO:19:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                #21                CGGA G                                                     - (2) INFORMATION FOR SEQ ID NO:20:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 18 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                #  18              TC                                                         - (2) INFORMATION FOR SEQ ID NO:21:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                # 20               TCTC                                                       - (2) INFORMATION FOR SEQ ID NO:22:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                # 20               CCTC                                                       - (2) INFORMATION FOR SEQ ID NO:23:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                # 19               AGG                                                        - (2) INFORMATION FOR SEQ ID NO:24:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                #21                TCAG G                                                     - (2) INFORMATION FOR SEQ ID NO:25:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                #                 22AGT TC                                                    - (2) INFORMATION FOR SEQ ID NO:26:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                # 19               CCA                                                        - (2) INFORMATION FOR SEQ ID NO:27:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 17 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                #   17             G                                                          - (2) INFORMATION FOR SEQ ID NO:28:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 17 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                #   17             G                                                          - (2) INFORMATION FOR SEQ ID NO:29:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 14 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                #     14                                                                      - (2) INFORMATION FOR SEQ ID NO:30:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                # 19               AGG                                                        - (2) INFORMATION FOR SEQ ID NO:31:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 15 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                #    15                                                                       - (2) INFORMATION FOR SEQ ID NO:32:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                # 20               AGCC                                                       - (2) INFORMATION FOR SEQ ID NO:33:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 18 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                #  18              AG                                                         - (2) INFORMATION FOR SEQ ID NO:34:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 16 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                #    16                                                                       - (2) INFORMATION FOR SEQ ID NO:35:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                #              26  GAAA ATGGGG                                                - (2) INFORMATION FOR SEQ ID NO:36:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                #             27   GGCG CCAGGGG                                               - (2) INFORMATION FOR SEQ ID NO:37:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                #               25 CGCC AGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:38:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 31 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                #          31      TCTG AAAATGGATA A                                          - (2) INFORMATION FOR SEQ ID NO:39:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 30 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                #           30     CTGA AAATGGATAA                                            - (2) INFORMATION FOR SEQ ID NO:40:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 29 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                #            29    ATTC TGAAAGGGG                                             - (2) INFORMATION FOR SEQ ID NO:41:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 28 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                #             28   ATTC TGAAAGGG                                              - (2) INFORMATION FOR SEQ ID NO:42:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 29 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                #            29    CTTC TTCCTGGGG                                             - (2) INFORMATION FOR SEQ ID NO:43:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 30 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                #           30     CTTC TTCCTGGGGG                                            - (2) INFORMATION FOR SEQ ID NO:44:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 31 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                #          31      CTTC TTCCTGGGGG G                                          - (2) INFORMATION FOR SEQ ID NO:45:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 30 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                #           30     CCTG CCATAGGGGG                                            - (2) INFORMATION FOR SEQ ID NO:46:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 29 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                #            29    CCTG CCATAGGGG                                             - (2) INFORMATION FOR SEQ ID NO:47:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                #                 22TCG GG                                                    - (2) INFORMATION FOR SEQ ID NO:48:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                #21                GGTC G                                                     - (2) INFORMATION FOR SEQ ID NO:49:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                #                 22GGG GG                                                    - (2) INFORMATION FOR SEQ ID NO:50:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                #21                AGGG G                                                     - (2) INFORMATION FOR SEQ ID NO:51:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                # 20               GGGG                                                       - (2) INFORMATION FOR SEQ ID NO:52:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                # 19               GGG                                                        - (2) INFORMATION FOR SEQ ID NO:53:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                #               25 AGGA GGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:54:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                #              26  GAGG AGGGGG                                                - (2) INFORMATION FOR SEQ ID NO:55:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                #                23AGCG GGG                                                   - (2) INFORMATION FOR SEQ ID NO:56:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                #                 22CGG GG                                                    - (2) INFORMATION FOR SEQ ID NO:57:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                #                23GGCA CAC                                                   - (2) INFORMATION FOR SEQ ID NO:58:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                #             27   GGCA CACGGGG                                               - (2) INFORMATION FOR SEQ ID NO:59:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                # 19               CAT                                                        - (2) INFORMATION FOR SEQ ID NO:60:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                #                23CATG GGG                                                   - (2) INFORMATION FOR SEQ ID NO:61:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                #21                CGGG C                                                     - (2) INFORMATION FOR SEQ ID NO:62:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                #               25 CGGG CGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:63:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                #21                CGGG G                                                     - (2) INFORMATION FOR SEQ ID NO:64:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                #                24GTCG AAAG                                                  - (2) INFORMATION FOR SEQ ID NO:65:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                #             27   GTCG AAAGGGG                                               - (2) INFORMATION FOR SEQ ID NO:66:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                #              26  TCGA AAGGGG                                                - (2) INFORMATION FOR SEQ ID NO:67:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                #               25 GAAA GGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:68:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                #              26  TCGA AGGGGG                                                - (2) INFORMATION FOR SEQ ID NO:69:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                #21                GGGG G                                                     - (2) INFORMATION FOR SEQ ID NO:70:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                #                 22GGG GG                                                    - (2) INFORMATION FOR SEQ ID NO:71:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                #               25 CCAC GGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:72:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                #                24CCAC GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:73:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 19 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                # 19               CGC                                                        - (2) INFORMATION FOR SEQ ID NO:74:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                #                23CGCG GGG                                                   - (2) INFORMATION FOR SEQ ID NO:75:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                #                24GCGC GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:76:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                #                24GCGC GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:77:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                #              26  TCCT CCGGGG                                                - (2) INFORMATION FOR SEQ ID NO:78:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                #21                TGAG G                                                     - (2) INFORMATION FOR SEQ ID NO:79:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                #                24TGAG GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:80:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                #                23TTCG GAG                                                   - (2) INFORMATION FOR SEQ ID NO:81:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                #              26  TTCG GAGGGG                                                - (2) INFORMATION FOR SEQ ID NO:82:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 17 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                #   17             C                                                          - (2) INFORMATION FOR SEQ ID NO:83:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                #21                CGGG G                                                     - (2) INFORMATION FOR SEQ ID NO:84:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                #                 22CGG GG                                                    - (2) INFORMATION FOR SEQ ID NO:85:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 21 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                #21                ATAT C                                                     - (2) INFORMATION FOR SEQ ID NO:86:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                #               25 ATAT CGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:87:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                #                24CCCC TCTC                                                  - (2) INFORMATION FOR SEQ ID NO:88:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                #                24TCTC GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:89:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 28 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                #             28   CCCC TCTCGGGG                                              - (2) INFORMATION FOR SEQ ID NO:90:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                #             27   TCCC CTCGGGG                                               - (2) INFORMATION FOR SEQ ID NO:91:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                #              26  CCCC TCGGGG                                                - (2) INFORMATION FOR SEQ ID NO:92:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                #                 22CGA GG                                                    - (2) INFORMATION FOR SEQ ID NO:93:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                #                24GCGA GGGG                                                  - (2) INFORMATION FOR SEQ ID NO:94:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 25 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                #               25 GCGA GGGGG                                                 - (2) INFORMATION FOR SEQ ID NO:95:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                #                23TCAG GGG                                                   - (2) INFORMATION FOR SEQ ID NO:96:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                #             27   TGTC TCAGGGG                                               - (2) INFORMATION FOR SEQ ID NO:97:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                #              26  TCAA GAGTTC                                                - (2) INFORMATION FOR SEQ ID NO:98:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                #              26  GAGT TCGGGG                                                - (2) INFORMATION FOR SEQ ID NO:99:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 24 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                #                24TTGC AGCC                                                  - (2) INFORMATION FOR SEQ ID NO:100:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 28 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                               #             28   TTGC AGCCGGGG                                              - (2) INFORMATION FOR SEQ ID NO:101:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 29 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                               #            29    TTGC AGCCGGGGG                                             - (2) INFORMATION FOR SEQ ID NO:102:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 26 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                               #              26  TTGC AGGGGG                                                - (2) INFORMATION FOR SEQ ID NO:103:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 22 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                               #                 22GGG GG                                                    - (2) INFORMATION FOR SEQ ID NO:104:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 23 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                               #                23GAGG GGG                                                   - (2) INFORMATION FOR SEQ ID NO:105:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 20 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                               # 20               GGGG                                                       __________________________________________________________________________

What is claimed is:
 1. An oligonucleotide of the formula

    5'-(CAP)-(Oligo)-(CAP)-3'

where Oligo is a nucleotide sequence of from 10 to 40 nucleotides inlength complementary to a target sequence, and CAP is G_(m), where m isan integer of from zero to ten, and where the two CAPs which are presentin the formula (a) are not complementary to the target sequence of theOligo and (b) are defined independently of each other, wherein (i) m maynot be zero at both the 5' and 3' ends and (ii) m may not be the same atboth the 5' and 3' ends when one end of said nucleotide sequence isguanine.
 2. The oligonucleotide as claimed in claim 1, wherein m is fromtwo to six.
 3. The oligonucleotide as claimed in claim 2, wherein m isfrom three to five.
 4. The oligonucleotide as claimed in claim 3,wherein m is four.
 5. The oligonucleotide as claimed in claim 1, whereinsaid nucleotide sequence is complementary to a transcribed DNA region ora translated mRNA.
 6. The oligonucleotide as claimed in claim 1, whereinOligo is selected from the group of nucleotide sequences consisting of(SEQ ID NO:1-34, respectively):

    ACACCCAATTCTGAAAATGG,                                      (I)

    AGGTCCCTGTTCGGGCGCCA,                                      (II)

    GTCGACACCCAATTCTGAAAATGGATAA,                              (III)

    GCTATGTCGACACCCAATTCTGAAA,                                 (IV)

    GTCGCTGTCTCCGCTTCTTCTTCCTG,                                (V)

    GTCTCCGCTTCTTCTTCCTGCCATAGG,                               (VI)

    GCGGGGCTCCATGGGGGTCG,                                      (VII)

    CAGCTGCAACCCAGC,                                           (VIII)

    GGCTGCTGGAGCGGGGCACAC,                                     (IX)

    AACGTTGAGGGGCAT,                                           (X)

    GTGCCGGGGTCTTCGGGC,                                        (XI)

    GGACAACATCATGGTCGAAAG,                                     (XII)

    CCCGAGAACATCATGGTCGAAG,                                    (XIII)

    GGGGAAAGCCCGGCAAGGGG,                                      (XIV)

    CACCCGCCTTGGCCTCCCAC,                                      (XV)

    GGGACTCCGGCGCAGCGC,                                        (XVI)

    GGCAAACTTTCTTTTCCTCC,                                      (XVII)

    GGGAAGGAGGAGGATGAGG,                                       (XVIII)

    GGCAGTCATCCAGCTTCGGAG,                                     (XIX)

    GCAGTAAGCATCCATATC,                                        (XX)

    CCCCCACCACTTCCCCTCTC,                                      (XXI)

    CTCCCCCACCACTTCCCCTC,                                      (XXII)

    GCTGGGAGCCATAGCGAGG,                                       (XXIII)

    ACTGCTGCCTCTTGTCTCAGG,                                     (XXIV)

    CAATCAATGACTTCAAGAGTTC,                                    (XXV)

    GGTCCCTGTTCGGGCGCCA,                                       (XXVI)

    GTGCCGGGGTCTTCGGG,                                         (XXVII)

    GGAGGATGCTGAGGAGG,                                         (XXVIII)

    GGAGGATGCTGAGG,                                            (XXIX)

    CAGGAGGATGCTGAGGAGG,                                       (XXX)

    GGCTGCCATGGTCCC,                                           (XXXI)

    TCATGGTGTCCTTTGCAGCC,                                      (XXXII)

    TCATGGTGTCCTTTGCAG,                                        (XXXIII)

and

    AAGTTCATGGTTTCGG                                           (XXXIV).


7. The oligonucleotide as claimed in claim 1, comprising the sequence(SEQ ID NO:30) C*A*GGAGGAT*GC*T*GAGGA*G*G, wherein * represents aphosphorothioate bridge.
 8. The oligonucleotide as claimed in claim 1,comprising the sequence (SEQ ID NO:53) G*G*G*CAGGAGGAT*GC*T*GAGGAGG*G*G*G, wherein * represents a phosphorothioate bridge.
 9. Theoligonucleotide as claimed in claim 1, comprising the sequence (SEQ IDNO:105) PY-G*G*G*GGAGGAT*GC*TGAGG*G*G*G, wherein * represents aphosphorothioate bridge and PY is pyrene.
 10. The oligonucleotide asclaimed in claim 1, comprising the sequence (SEQ ID NO:104)G*G*G*GGAGGAT*GC*T*GAGGAGG*G*G*G, wherein * represents aphosphorothioate bridge.
 11. The oligonucleotide as claimed in claim 1,comprising the sequence (SEQ ID NO:51) G*G*G*GGAGGAT*GC*TGAGG*G*G*G. 12.The oligonucleotide as claimed in claim 1, wherein at least onenucleotide is modified.
 13. The oligonucleotide as claimed in claim 12,wherein from 2 to 10 non-terminal pyrimidine nucleosides are modified.14. The oligonucleotide an claimed in claim 12, wherein theoligonucleotide is modified at the 5'-terminal and/or 3'-terminalnucleotide(s).
 15. The oligonucleotide as claimed in claim 12, whereinat least one group of 1 to 4 nucleotides which are connected to eachother is not modified.
 16. The oligonucleotide as claimed in claim 12,wherein the modification is selected from the group consisting of:(a)complete or partial replacement of the 3' and/or 5' phosphoric diesterbridges, (b) complete or partial replacement of the 3' and/or 5'phosphoric diester bridges by dephospho bridges, (c) complete or partialreplacement of the sugar phosphate backbone, (d) complete or partialreplacement of the β-D-2'-deoxyribose units, and (e) complete or partialreplacement of the natural nucleoside bases.
 17. The oligonucleotide asclaimed in claim 16, wherein the modification is selected from the groupconsisting of(a) complete or partial replacement of the 3' and/or 5'phosphoric diester bridges by a member selected from the groupconsisting of phosphorothioate, phosphorodithioate, (NR¹PR²)-phosphoramidate, boranophosphate, phosphate-(C₁ -C₂₁)-O-alkylester, phosphate[(C₆ -C₁₂)aryl-(C₁ -C₂₁)-O-alkyl] ester,2,2,2-trichlorodimethylethylphosphonate, (C₁ -C₈)-alkylphosphonate and(C₆ -C₁₂)-arylphosphonate bridge, where R¹ and R₂ are, independently ofeach other, hydrogen, or (C₁ -C₈)-alkyl, (C₆ -C₂₀)-aryl, (C₆-C₁₄)-aryl-(C₁ -C₈)-alkyl or --(CH₂)_(c) --[NH(CH₂)_(c) ]_(d) NR³ R³, inwhich c is an integer of from 2 to 6, d is an integer of from 0 to 6,the R³ groups are, independently of each other, hydrogen, (C₁ -C₆) alkylor (C₁ -C₄)-alkoxy-(C₁ -C₆)-alkyl, R¹ and R² are independently hydrogen,(C₁ -C₈)-alkyl or methoxyethyl, or R¹ and R² together with the nitrogenatom carrying them, form a 5-6-membered heterocyclic ring which canadditionally contain a further O, S, or N hetero atom; (b) replacementof one, two or three of the 3' and/or 5' phosphoric diester bridge(s) bydephospho bridge(s) selected from the group consisting of formacetal,3'-thioformacetal, methylhydroxyl-amine, oxime,methylenedimethylhydrazo, dimethylenesulfone and silyl; (c) complete orpartial replacement of the sugar phosphate backbone by amorpholinonucleotide oligomer; (d) replacement of one, two, or three ofthe β-D-2'-deoxyribose units at the 5' end and/or at the 3' end by amember selected from the group consisting of α-D-2'-deoxyritose,L-2'-deoxyribose, 2'-F-2'deoxyribose, 2'-O-(C₁ -C₆)alkyl-ribose,2'-O-(C₂ -C₆)-alkenyl-ribose, 2'-NH₂ -2'-deoxyribose, β-D-xylofuranose,α-arabinofuranose, 2,4-dideoxy-β-D-erythrohexopyranose, and carbocyclic,open-chain and bicyclo sugar analogs; and (e) complete or partialreplacement of the natural nucleoside bases other than the naturalnucleoside bases of the CAP regions by a member selected from the groupconsisting of 5-(hydroxymethyl)uracil, 5-aminouracil, pseudouracil,dihydrouracil, 5-(C₁ -C₆)-alkyl-uracil, 5-(C₂ -C₆)-alkenyl-uracil, 5-(C₂-C₆)-alkynyluracil, 5-(C₁ -C₆)-alkyl-cytosine, 5-(C₂-C₆)alkenyl-cytosine, 5-(C₂ -C₆)-alkynyl-cytosine, 5-fluorouracil,5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil and5-bromocytosine.
 18. The oligonucleotide as claimed in claim 17, whereinin definition a) the modification is selected from the group consistingof a phosphorothioate, phosphorodithioate, (NR¹ R²)-phosphoramidate,phosphate-O-methyl ester, phosphate-O-ethyl ester, phosphate-O-isopropylester, methylphosphonate and phenylphosphonate bridge.
 19. Theoligonucleotide as claimed in claim 18, wherein the modification isselected from the group consisting of a phosphorothioate,phosphorodithioate and methylphosphonate bridge.
 20. The oligonucleotideas claimed in claim 19, wherein the modification is a phosphorothioatebridge.
 21. The oligonucleotide as claimed in claim 17, wherein indefinition a) R¹ and R² independently represent hydrogen, (C₁ -C₄)-alkylor methoxyethyl.
 22. The oligonucleotide as claimed in claim 17, whereinin definition b) the modification is selected from the group consistingof formacetal and 3'-thioformacetals.
 23. The oligonucleotide as claimedin claim 17, wherein in definition b) the one, two or three phosphoricdiester bridges are replaced at the 5' end and at the 3' end.
 24. Theoligonucleotide as claimed in claim 23, wherein the phosphoric diesterbridges are replaced at the pyrimidine positions.
 25. Theoligonucleotide as claimed in claim 17, wherein in definition d) themodification is selected from the group consisting of2'-F-2'-deoxyribose, 2'-O-(C₁ -C₆)alkylribose, 2'-O-(C₂-C₆)alkenyl-ribose and 2'-NH₂ -2'-deoxyribose.
 26. The oligonucleotideas claimed in claim 25, wherein the modification is selected from thegroup consisting of 2'-F-2'-deoxyribose, 2'-O-(C₁ -C₄)alkylribose,2'-O-(C₂ -C₄)alkenyl-ribose and 2'-NH₂ -2'-deoxyribose.
 27. Theoligonucleotide as claimed in claim 26, wherein the modification isselected from the group consisting of 2'-O-methyl-, 2'-O-allyl-, and2'-O-butylribose.
 28. The oligonucleotide as claimed in claim 17,wherein in definition d) the one, two or three ribose units are replacedat the 5' end and at the 3' end.
 29. The oligonucleotide as claimed inclaim 28, wherein the ribose units are replaced at the pyrimidinepositions.
 30. The oligonucleotide as claimed in claim 17, wherein indefinition e) the modification is selected from the group consisting ofa 5-(C₁ -C₆)-alkyluracil, 5-(C₂ -C₆)-alkenyl-uracil, 5-(C₂-C₆)alkynyl-uracil, 5-(C₁ -C₆)-alkyl-cytosine, 5-(C₂-C₆)-alkenyl-cytosine, 5-(C₂ -C₆)-alkynylcytosine, 5-fluorouracil,5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, and5-bromocytosine.
 31. The oligonucleotide as claimed in claim 30, whereinthe modification is selected from the group consisting of 5-(C₃-C₆)-alkyl-uracil, 5-(C₂ -C₆)-alkenyl-uracil, 5-(C₂ -C₆)-alkynyl-uracil,5-(C₁ -C₆)alkyl-cytosine, 5-(C₂ -C₆)-alkenyl-cytosine, and 5-(C₂-C₆)-alkynyl-cytosine.
 32. The oligonucleotide as claimed in claim 31,wherein the modification is selected from the group consisting of5-pentynylcytosine, 5-hexynyluracil, and 5-hexynylcytosine.
 33. Theoligonucleotide as claimed in claim 1, which is linked, at the 5' endand/or the 3' end to a molecule selected from the group consisting ofpolylysine, intercalators, fluorescent compounds, crosslinkers,lipophilic molecules, lipids, steroids, vitamins, polyethylene glycol,oligoethylene glycol, (C₁₂ -C₁₈)-alkyl-phosphate diesters, and --O--CH₂--CH(OH)O--(C₁₂ -C₁₈)-alkyl.
 34. The oligonucleotide as claimed in claim33 wherein the intercalator is selected from the group consisting ofpyrene, acridine, phenazine and phenanthridine.
 35. The oligonucleotideas claimed in claim 33 wherein the fluorescent compound is fluorescein.36. The oligonucleotide as claimed in claim 33 wherein the crosslinkeris selected from the group consisting of psoralene and azidoproflavin.37. The oligonucleotide as claimed in claim 33 wherein the lipophilicmolecule is (C₁₂ -C₂₀)-alkyl.
 38. The oligonucleotide as claimed inclaim 33 wherein the lipid is 1,2-dihexadecyl-rac-glycerol.
 39. Theoligonucleotide as claimed in claim 33 wherein the steroid ischolesterol or testosterone.
 40. The oligonucleotide as claimed in claim33 wherein the vitamin is vitamin E.
 41. The oligonucleotide as claimedin claim 1, which contains a 3'--3' inversion and/or a 5'--5' inversionat the 5' end and/or 3' end.
 42. A process for preparing anoligonucleotide as claimed in claim 1, which comprises synthesizing thenucleotide sequence chemically from individual nucleotides.
 43. Apharmaceutical composition comprising one or more oligonucleotides asclaimed in claim 1 and a physiologically acceptable carrier.
 44. In amethod of treating a disorder selected from the group consisting ofcancer and restenosis comprising administering as a therapeutic agent anoligonucleotide complementary to an RNA associated with said disorder,the improvement comprising administering to a subject in need thereof aneffective amount of the composition of claim
 43. 45. A method ofdiagnosing a disorder selected from the group consisting of cancer,restenosis, a disease caused by a virus, a disease affected by integrinsor cell-cell adhesion receptors and a disease triggered by diffusiblefactors, comprising taking a sample from a subject, adding anoligonucleotide as claimed in claim 1 to the sample, and detectingwhether or not the oligonucleotide hybridizes to a DNA sequencecontained in the sample.
 46. The oligonucleotide as claimed in claim 1,wherein m is not zero at the 5'-end.